CAJ | 학술논문

构建双基因共表达腺病毒载体转染关节炎模型大鼠的意义
구건쌍기인공표체선병독재체전염관절염모형대서적의의
Transfection of double gene co-expressing adenovirus vector into arthritis rats

万方数据

中国组织工程研究中國組織工程研究 중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2015年 18期 2825-2830 ,共6页

樊萍%何岚%郝志明%蒲丹%吕晓虹%孙怡宁%胡楠%王研华%丁小明%李杨%薛武军

번평%하람%학지명%포단%려효홍%손이저%호남%왕연화%정소명%리양%설무군

实验动物%骨及关节损伤模型%关节炎%类风湿%CD40L%IκBα%核因子κB%共刺激信号%炎症%滑膜%国家自然科学基金實驗動物%骨及關節損傷模型%關節炎%類風濕%CD40L%IκBα%覈因子κB%共刺激信號%炎癥%滑膜%國傢自然科學基金
실험동물%골급관절손상모형%관절염%류풍습%CD40L%IκBα%핵인자κB%공자격신호%염증%활막%국가자연과학기금
Arthritis%Adenoviridae Infections%Lymphocytes%T-Lymphocytes

背景：淋巴细胞异常激活以及核因子κB依赖非特异性的炎症反应是类风湿关节炎关节组织炎症损害的两个重要方面。共刺激信号CD40/CD40L是T细胞识别、活化中最重要的共刺激分子。IκBα有效的抑制核因子κB的途径，可以在中心环节上抑制炎症反应的产生，抑制炎症因子滑膜组织的损害。 　　目的：采用 CD40LIg-IRES2-IκBα双基因表达的腺病毒载体转染到关节炎动物模型，探索双基因共表达腺病毒载体转染对免疫性关节炎的治疗效果。 　　方法：构建pAdCD40LIg-IRES2-IκBα双基因共表达腺病毒载体。采用含1 g/LⅡ型胶原蛋的完全弗氏佐剂皮下多点注射构建关节炎Wistar大鼠模型。将20只构建成功的关节炎大鼠模型分为未处理组和转染组，分别给予2组大鼠四肢末端关节腔注射生理盐水和pAdCD40LIg-IRES2-IκBα腺病毒载体。 　　结果与结论：转染14 d后，与未处理组相比，转染组大鼠关节炎评分、关节液中淋巴细胞CD40L表达水平、关节滑膜组织中核因子κB p65表达水平、关节液内白细胞介素2、白细胞介素6、肿瘤坏死因子α、间质金属蛋白酶3和间质金属蛋白酶9的水平降低。提示共表达CD40LIg和IκBα腺病毒载体局部转染可有效抑制关节炎大鼠关节炎症状，降低关节腔内炎性细胞因子及炎性分子在滑膜组织中的表达，取得良好的治疗效果。
배경：림파세포이상격활이급핵인자κB의뢰비특이성적염증반응시류풍습관절염관절조직염증손해적량개중요방면。공자격신호CD40/CD40L시T세포식별、활화중최중요적공자격분자。IκBα유효적억제핵인자κB적도경，가이재중심배절상억제염증반응적산생，억제염증인자활막조직적손해。 　　목적：채용 CD40LIg-IRES2-IκBα쌍기인표체적선병독재체전염도관절염동물모형，탐색쌍기인공표체선병독재체전염대면역성관절염적치료효과。 　　방법：구건pAdCD40LIg-IRES2-IκBα쌍기인공표체선병독재체。채용함1 g/LⅡ형효원단적완전불씨좌제피하다점주사구건관절염Wistar대서모형。장20지구건성공적관절염대서모형분위미처리조화전염조，분별급여2조대서사지말단관절강주사생리염수화pAdCD40LIg-IRES2-IκBα선병독재체。 　　결과여결론：전염14 d후，여미처리조상비，전염조대서관절염평분、관절액중림파세포CD40L표체수평、관절활막조직중핵인자κB p65표체수평、관절액내백세포개소2、백세포개소6、종류배사인자α、간질금속단백매3화간질금속단백매9적수평강저。제시공표체CD40LIg화IκBα선병독재체국부전염가유효억제관절염대서관절염증상，강저관절강내염성세포인자급염성분자재활막조직중적표체，취득량호적치료효과。
BACKGROUND:Abnormal activation of lymphocytes and nuclear factorκB-dependent non-specific inflammation are two major manifestations of joint damage in rheumatoid arthritis. Co-stimulatory signal CD40/CD40L is the dominant co-stimulatory factor in the recognition and activation of T cel s. IκBαeffectively inhibits nuclear factorκB pathway, prevent the inflammation in the central link, and suppress the damage caused by inflammatory factor in the synovial tissue. OBJECTIVE:To investigate the therapeutic effect of double gene co-expressing adenovirus vector on arthritis based on an arthritis model rat transfected by CD40LIg-IRES2-IκBαco-expressing adenovirus vector. METHODS:The pAdCD40LIg-IRES2-IκBαco-expressing adenovirus vector was established. Arthritic model was established through multi-subcutaneous injections of complete Freund's adjuvant of type col agen II (1 g/L) into Wistar rats. Then 20 arthritic rats were divided into two groups:untreated group and transfection group, receiving an injection of saline and pAdCD40LIg-IRES2-IκBαadenovirus vector to distal joint cavity of limbs, respectively. RESULTS AND CONCLUSION:At 14 days post-transfection, compared with the untreated group, the mean arthritis index score, the CD40L expression of lymphocytes in synovial fluid, the nuclear factor-κB p65 expression in synovial tissue, and levels of interleukin-2, interleukin-6, tumor necrosis factor-α, matrix metal oproteinase-3 and matrix metal oproteinase-9 in synovial fluid of rats in transfection group were significantly lower than those in untreated group. Focal transfection of the CD40LIg-IκBαco-expression adenovirus vector can effectively inhibit arthritic symptoms, and reduce the expressions of inflammatory cytokine in synovial fluid and inflammatory molecule in synovial tissue of arthritic rats, which shows good therapeutic effect.